PROTEOMICS SHARED RESOURCE PROJECT SUMMARY The Mayo Clinic Cancer Center (MCCC) Proteomics Shared Resource (PRO) is a full-service facility that provides essential services in peptide synthesis, protein analysis, protein and peptide separation, mass spectrometry, proteomics applications, and expertise in project planning, sample preparation, data acquisition and analysis. The PRO occupies a total of 4920 sq. ft. on the 3rd floor of the Medical Sciences Building, and is located at the Mayo Clinic in Rochester (MCR) site. The 8 PRO staff members have broad knowledge and expertise in the services provided. The PRO Director, Dr. Daniel J. McCormick, provides visionary direction, planning, recruitment, and executive decisions regarding all operations, staffing, and services. Working with institutional committees and centers for over 33 years, Dr. McCormick has been able to provide scientific direction and support for high-end core instrumentation, and new technologies in protein chemistry, proteomics and mass spectrometry (MS) to support Programs of MCCC members in cancer research. In the previous funded period, 6 new service-lines (e.g., label-free differential proteomics, expanded phosphoproteomics by SCX fractionation and TiO2 enrichment, proteomics analysis of FFPE and Frozen tissue, high/low pH LC sample fractionation, de novo MS/MS protein sequencing, and MS analysis of complex glycans) were developed to support the protein analysis needs of MCCC members. Scientific achievements include: identification of key proteins that interact with TAT2 protein in myeloid cancers; development of agents that inhibit TGF action (via SMAD signaling) and prevent glioblastoma migration in PDX cell lines; identification of key phosphoproteins (e.g., CXCR4) downregulated in Alisertib-treated human breast adenocarcinoma; mapping sites of phosphorylation in SOX2, a transcriptional activator of ?stemness? phenotype in lung squamous cell carcinoma; synthesis of oncoprotein antigens for in vitro activation of nave T-cells in adoptive T-cell therapies to treat cancer (e.g., pancreatic); and differential analysis of proteins activated by SPINK1 in ovarian cancer proliferation. PRO utilization by MCCC members has been strong with a total average usage of 51% by 99 members (72 peer-reviewed) in the current funded period (2013-2017). The PRO is used by MCCC members from all 3 sites, including 12 members from Mayo Clinic in Florida and 5 members from Mayo Clinic in Arizona. The PRO provides free consultation, project design, and data interpretation to MCCC members, and provides its services on fee-for-service basis; standardized forms for project and service requests are on the resources' internal web site. Comparison of rates for many services of the PRO to external cores and vendors, was completed in 2017 and indicated that core rates were highly competitive. Future directions of the PRO include: deep sequencing proteomics with advanced MS instrumentation and data-independent acquisition (DIA) methods; top-down MS protein sequencing; ion mobility separation (IMS) mass spectrometry; proteogenomics integration of genomics/proteomics data sets, and MS analysis of glycoproteins.